Miyaji K, Kaneko S, Ishikawa H, Aoyagi T, Hayakawa K, Hata M, Oohashi M, Izawa A, Murai M
Department of Urology, Ichikawa General Hospital, Tokyo Dental College, Chiba, Japan.
Arch Androl. 2001 Mar-Apr;46(2):127-34.
In previous conventional reports sperm and seminal plasma were not well separated and therefore it was highly probable that isoform (CK-BB) of seminal plasma was determined together with that of sperm. The authors developed a method to purify sperm specimen free from seminal plasma. Human sperm was purified by means of the continuous-step density gradient centrifugation with an inner column and the subsequent modified swim-up procedure. CK isoforms in the purified sperm and the seminal plasma were analyzed by ion exchange chromatography and electrophoresis. The purified sperm contained mitochondrial CK-MiMi as well as cytosolic CK, of which isoform was almost occupied by CK-MM but CK-BB was not detected. On the other hand, CK isoform in the seminal plasma was detected as CK-BB alone. CK activity in human sperm and seminal plasma were 2.0 +/- 0.7 x 10(-6) IU/10(6) sperm and 661 +/- 360 IU/L, respectively. CK activity in the seminal plasma gave no correlation with the semen parameters.
在以往的传统报告中,精子和精浆没有得到很好的分离,因此很有可能精浆的同工型(CK-BB)与精子的同工型一起被测定。作者开发了一种从精浆中纯化精子标本的方法。通过使用内柱的连续梯度密度离心法和随后的改良上游法对人类精子进行纯化。通过离子交换色谱法和电泳法分析纯化精子和精浆中的CK同工型。纯化的精子含有线粒体CK-MiMi以及胞质CK,其同工型几乎全部为CK-MM,但未检测到CK-BB。另一方面,精浆中的CK同工型仅检测为CK-BB。人类精子和精浆中的CK活性分别为2.0±0.7×10⁻⁶ IU/10⁶精子和661±360 IU/L。精浆中的CK活性与精液参数无关。